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Title: Comparative molecular analysis of two asparaginyl endopeptidases and encoding genes from Fasciola gigantica
Authors: Poom Adisakwattana
Vithoon Viyanant
Wanpen Chaicumpa
Suksiri Vichasri-Grams
Annemarie Hofmann
Günter Korge
Prasert Sobhon
Rudi Grams
Thammasat University
Mahidol University
Freie Universitat Berlin
Keywords: Biochemistry, Genetics and Molecular Biology;Immunology and Microbiology
Issue Date: 1-Dec-2007
Citation: Molecular and Biochemical Parasitology. Vol.156, No.2 (2007), 102-116
Abstract: In this study we describe the first cysteine proteinases of the MEROPS Clan CD family C13 in Fasciola gigantica. Family C13 contains asparaginyl endopeptidases and glycosylphosphatidylinositol-anchor transamidases and is also called the legumain family due to the discovery of the first asparaginyl endopeptidase in a legume. The cDNAs encoding two asparaginyl endopeptidases, FgLGMN-1 and FgLGMN-2, were cloned and used for the analysis of nucleic acid and protein properties. The deduced amino acid sequences showed 47.4% identity to each other and from 42.2 to 51.1% identity to homologs of other trematode species. The catalytic site residues histidine, cysteine and preceding hydrophobic residues, characteristic for the cysteine proteinase families C11, C13, C14, and C25, were found conserved. Northern and reverse transcription PCR analyses demonstrated that the transcriptional products are present in metacercariae, juveniles and adults. RNA in situ hybridization and immunohistochemistry revealed that RNA and protein products of the two genes are specifically expressed in the intestinal epithelium of juveniles and adults. Immune sera of mice infected with F. gigantica reacted with immunoblotted, bacterially expressed recombinant proteins starting 4 weeks after infection. Polyclonal antisera raised against the recombinant proteins detected 40 and 30 kDa antigens, respectively in crude worm protein extracts but not in the excretion-secretion products of adult parasites. Likewise, legumain-specific activity was found in crude worm protein extracts but not in excretion-secretion products. This study elucidates the molecular characteristics of these proteins in F. gigantica and demonstrates differences in the biology between Fasciola and Schistosoma which may prove useful for the development of vaccines against fasciolosis in domestic livestock. © 2007 Elsevier B.V. All rights reserved.
ISSN: 01666851
Appears in Collections:Scopus 2006-2010

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